Kinetics of UV-induced changes in deoxynucleoside triphosphate pools in Chinese hamster ovary cells and their effect on measurements of DNA synthesis

Measurements of dNTP pools following exposure of Chinese hamster ovary cells to ultraviolet radiation reveals a rapid accumulation of cellular dTTP and a rapid loss of cellular dCTP. Exposure to 3-, 10- or 20 Jm-2 results in a 3-, 4- or 5.4-fold increase in cellular dTTP, respectively, within the first 10 min after exposure. dTTP levels then decrease noticeably, approaching the control value 3 to 5 hr later. In contrast, dCTP levels decrease rapidly within 10 min after exposure, ultimately to 1/10 that observed in the unirradiated control population. Recovery to normal dCTP levels is slow, taking in excess of 12 hr. No change in dATP is observed for 1-2 hr; subsequently, a moderate decrease in dATP levels occurs which is then followed by recovery, beginning 8 hr after irradiation. These results contrast with changes in dNTP pools observed in Chinese hamster V-79 cells exposed to mutagens. Measurements of rates of DNA synthesis by pulse-labeling cells with [3H]thymidine are also apparently affected by UV-induced transient deviations in the endogenous radiospecific activity of the labeled precursor.     

A relationship between L-serine degradation and methionine biosynthesis in Escherichia coli K12

While wild-type Escherichia coli K12 cannot grow with L-serine as carbon source, two types of mutants with altered methionine metabolism can. The first type, metJ mutants, in which the methionine biosynthetic enzymes are expressed constitutively, are able to grow with L-serine as carbon source. Furthermore, a plasmid carrying the metC gene confers ability to grow on L-serine. These observations suggest that in these mutants, L-serine deamination may be a result of a side-reaction of the metC gene product, cystathionine beta-lyase. The second type is exemplified by two newly isolated strains carrying mutations mapping between 89.6 and 90 min. These mutants use L-serine as carbon source, and also require methionine for growth with glucose at 37 degrees C and above. The phenotypes of the new mutants resemble those of both met and his constitutive mutants in some respects, but have been differentiated from both of them.     

The respiratory burst response to Histoplasma capsulatum by human neutrophils. Evidence for intracellular trapping of superoxide anion

Human neutrophils (PMN) have received little attention as to the role they play in host defense against Histoplasma capsulatum (Hc). We have characterized the binding and phagocytosis of Hc yeasts by human PMN and quantified the PMN respiratory burst in response to this organism. mAb specific for CD11a, CD11b, and CD11c all partially blocked the attachment of unopsonized yeasts to PMN; a mAb to CD18 inhibited attachment by greater than 90%. Thus, human PMN recognize and bind Hc yeasts via CD18 adhesion receptors as has been found for human cultured macrophages and alveolar macrophages. Unopsonized yeasts were phagocytosed by PMN, but phagocytosis was increased markedly by heat-labile and heat-stable serum opsonins. These opsonins promoted enhanced phagocytosis of yeasts by increasing the attachment of Hc yeasts to the PMN membrane. Phagocytosis of viable or heat-killed Hc yeasts by PMN did not induce the secretion of superoxide anion (O2-) as quantified by the reduction of cytochrome c. O2- was not detected when yeasts were opsonized in normal serum or immune serum, or at a ratio of yeasts to PMN of up to a 100:1. However, phagocytosis of opsonized yeasts by PMN did not prevent them from subsequently releasing O2- after further incubation with opsonized zymosan or PMA. Opsonized Hc yeasts clearly stimulated the PMN respiratory burst as quantified by intracellular reduction of nitroblue tetrazolium, reduction of cytochrome c in the presence of cytochalasin D, oxygen consumption, luminol-enhanced and nonenhanced chemiluminescence, and H2O2 production. These data suggest that phagocytosis of Hc yeasts by PMN is associated with intracellular entrapment of O2- that is not detectable by reduction of extracellular cytochrome c.     

Rapid loss of phosphorus from dying ryegrass roots: the chemical components involved

Summary Lolium perenne was grown in solution culture with either ample or deficient phosphate supply (high-P and low-P plants). The concentration in the roots of phosphorus as water-soluble compounds, phospholipid and insoluble residue was measured. A supplementary experiment showed that the concentration of each component in the roots of low-P plants was similar to that in plants grown in P-deficient soil. The time-course of the decline of each P component was determined in roots detached from the shoot and left hanging in solution. During the three weeks residue P concentration in the roots declined little. In contrast, both types of root lost about three-quarters of their lipid P in the first week. Low-P roots lost little of their water-soluble P. High-P roots contained much more water-soluble P and lost much of it during the first two weeks. By the end of three weeks their water-soluble P content was levelling out at a value similar to that in low-P roots, suggesting a non-labile pool. The rapid loss of lipid P from low-P roots comprised more than half of their total loss, and the possible ecological significance of this is discussed.     

Structure-function relationship of human spinal ligaments

A combination of light, scanning and transmission electron microscopy was used to investigate the morphology and ultrastructure of normal human spinal ligaments sampled from adult surgical specimens. The ligamenta flava consist mostly of dense elastic fibers, whereas the supraspinous and interspinous ligaments are preponderantly collagenous. In all ligaments, the collagen fascicles are characterized by a regular crimp structure. The inner collagen fibers of interspinous ligaments tend to be oriented parallel to the spinous processes while those of the peripheral layers run in postero-cranial direction. The presence of proteoglycan filaments is clearly demonstrated in all of the ligaments examined. They are mainly located at the d band of the collagen fibrils. These findings are discussed in relation to the function of the posterior ligamentous system. It is suggested that the interspinous ligaments are able to transmit tension from the thoracolumbar fascia to the spine. Finally, the spinal ligaments are thought to be involved in the control mechanism of the spine.